MagnEdit-interacting factors that recruit DNA-editing enzymes to single base targets

Jennifer L McCann, Daniel Salamango, Emily K. Law, William L. Brown, Reuben S. Harris

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Although CRISPR/Cas9 technology has created a renaissance in genome engineering, particularly for gene knockout generation, methods to introduce precise single base changes are also highly desirable. The covalent fusion of a DNA-editing enzyme such as APOBEC to a Cas9 nickase complex has heightened hopes for such precision genome engineering. However, current cytosine base editors are prone to undesirable off-target mutations, including, most frequently, target-adjacent mutations. Here, we report a method to “attract” the DNA deaminase, APOBEC3B, to a target cytosine base for specific editing with minimal damage to adjacent cytosine bases. The key to this system is fusing an APOBEC-interacting protein (not APOBEC itself) to Cas9n, which attracts nuclear APOBEC3B transiently to the target site for editing. Several APOBEC3B interactors were tested and one, hnRNPUL1, demonstrated proof-of-concept with successful C-to-T editing of episomal and chromosomal substrates and lower frequencies of target-adjacent events.

Original languageEnglish (US)
Article numberLSA.201900606
JournalLife science alliance
Volume3
Issue number4
DOIs
StatePublished - Feb 24 2020

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